Gastroenteritis Outbreaks Caused by Norovirus GII.17, Guangdong Province, China, 2014-2015.

In the past decade, the most prevalent norovirus genotype causing viral gastroenteritis outbreaks worldwide, including China, has been GII.4. In winter 2014-15, norovirus outbreaks in Guangdong, China, increased. Sequence analysis indicated that 82% of the outbreaks were caused by a norovirus GII.17 variant.

Elucidation of echovirus 30's origin and transmission during the 2012 aseptic meningitis outbreak in Guangdong, China, through continuing environmental surveillance.

An aseptic meningitis outbreak occurred in Luoding City of Guangdong, China, in 2012, and echovirus type 30 (ECHO30) was identified as the major causative pathogen. Environmental surveillance indicated that ECHO30 was detected in the sewage of a neighboring city, Guangzhou, from 2010 to 2012 and also in Luoding City sewage samples (6/43, 14%) collected after the outbreak. In order to track the potential origin of the outbreak viral strains, we sequenced the VP1 genes of 29 viral strains from clinical patients and environmental samples. Sequence alignments and phylogenetic analyses based on VP1 gene sequences revealed that virus strains isolated from the sewage of Guangzhou and Luoding cities matched well the clinical strains from the outbreak, with high nucleotide sequence similarity (98.5% to 100%) and similar cluster distribution. Five ECHO30 clinical strains were clustered with the Guangdong environmental strains but diverged from strains from other regions, suggesting that this subcluster of viruses most likely originated from the circulating virus in Guangdong rather than having been more recently imported from other regions. These findings underscore the importance of long-term, continuous environmental surveillance and genetic analysis to monitor circulating enteroviruses.

[Epidemiological characteristics of norovirus variant of GII.4 Sydney and the outbreaks caused by norovirus variant of GII.4 Sydney in Guangdong province, 2012-2014].

OBJECTIVE: To analyze epidemiological characteristics of norovirus variant of GII.4 Sydney from January 2012 to June 2014 in sentinel hospitals of Guangdong province, as well as the outbreaks caused by norovirus variant of GII.4 Sydney. METHODS: During January 2012 to June 2014, a total of 10 750 fecal samples were obtained from 22 hospitals of surveillance sites in Guangdong province. Those samples were sent to the local municipal CDCs for extracting and detecting norovirus nucleic acid. Then, all the positive samples were delivered to Guangdong provincial CDC that used Random Number Method to draw 855 positive samples for norovirus genotyping, and 690 samples were successfully sequenced. Chi-square tests were used to compare norovirus infection status of diarrhea cases in different age groups as well as during different periods. Epidemiological data of 13 outbreaks which were caused by norovirus variant of GII.4 Sydney from January 2012 to June 2014 were collected from the Public Health Emergency Management Information System of Guangdong Province, and the epidemiological characteristics were analyzed. RESULTS: The norovirus variant of GII.4 Sydney was first detected in August 2012 and the detection rate was 13/15 in November 2012. During November 2012 to January 2013 (period T1), the norovirus positive rate of each month was 23.8% (100/421), 15.9% (61/383) and 19.2% (95/495), respectively. During November 2013 to January 2014 (period T2), the norovirus positive rate of each month was 17.0% (90/529), 8.7% (37/426) and 11.2% (46/409), respectively which were significantly lower than that of period T1 (χ² alue was 6.65, 9.93 and 10.74. P value was 0.010, 0.002, and 0.001, respectively). In period T1, the norovirus positive rate of people ages 15 and older was 26.3% (143/543) and the rate of people under 15 was 14.9% (113/756) (χ² = 2.90, P < 0.001). In period T2, the norovirus positive rate of people ages 15 and older was 10.1% (52/516) and the rate of people under 15 (14.3% (121/848)) (χ²= 5.09, P = 0.024). The foodborne transmission was the infection source for ten of thirteen outbreaks. CONCLUSION: The norovirus variant of GII.4 Sydney was first detected in August 2012. The epidemic began to occur in the community since November 2012, and the strength of the epidemic declined 1 year later. The foodborne transmission was the main infection sources for the outbreaks caused by norovirus variant of GII.4 Sydney.

Prevalence of nonpolio enteroviruses in the sewage of Guangzhou city, China, from 2009 to 2012.

The human-pathogenic viruses in urban sewage have been extensively monitored to obtain information on circulating viruses in human communities. Enteroviruses (EVs) excreted by patients who present with diverse clinical syndromes can remain infectious in the environment for several weeks, and limited data on circulating environmental EVs are available. A 4-year (2009 to 2012) surveillance study was conducted to detect nonpolio enteroviruses (NPEVs) in the urban sewage of Guangzhou city, China. After the viruses in the sewage samples were concentrated and isolated, molecular identification was used to detect and type the NPEVs. During the 4-year study, 17 different NPEV serotypes were identified in the sewage of Guangzhou city. The most common serotypes were echovirus 11 (ECHO11), ECHO6, ECHO7, and ECHO12 and coxsackie group B viruses 5 (CVB5) and CVB3. The predominant serotypes were influenced by spatial and temporal factors and differed each year. CVB5 was commonly detected in 2009 and 2010 but was rarely isolated in 2011 and 2012. In contrast, CVB3 was not observed in 2009 and 2010 but was increasingly detected in 2011 and 2012. Our study provides an overview of the serotype distribution and circulation patterns of NPEVs in the sewage of Guangzhou, China. In the absence of a systematic EV disease surveillance system, the detection and characterization of sewage-borne NPEVs will help us better understand the changes in EV disease trends and the epidemic background of circulating EVs, which could help interpret the EV trends and warn of future outbreaks in this area.

Exploring the action mechanism of Jinxin oral liquid on asthma by network pharmacology, molecular docking, and microRNA recognition.

Using network pharmacology, molecular docking, and microRNA recognition, we have elucidated the mechanisms underlying the treatment of asthma by Jinxin oral liquid (JXOL). We began by identifying and normalizing the active compounds in JXOL through searches in the traditional Chinese medicine systems pharmacology database, SwissADME database, encyclopedia of traditional Chinese medicine database, HERB database, and PubChem. Subsequently, we gathered and standardized the targets of these active compounds from sources including the encyclopedia of traditional Chinese medicine database, similarity ensemble approach dataset, UniProt, and other databases. Disease targets were extracted from GeneCards, PharmGKB, OMIM, comparative toxicogenomics database, and DisGeNET. The intersection of targets between JXOL and asthma was determined using a Venn diagram. We visualized a Formula-Herb-Compound-Target-Disease network and a protein-protein interaction network using Cytoscape 3.9.0. Molecular docking studies were performed using Schrodinger software. To identify pathways related to asthma, we conducted gene ontology functional analysis and Kyoto encyclopedia of genes and genomes pathway enrichment analysis using Metascape. MicroRNAs regulating the hub genes were obtained from the miRTarBase database, and a network linking these targets and miRNAs was constructed. Finally, we found 88 bioactive components in JXOL and 218 common targets with asthma. Molecular docking showed JXOL key compounds strongly bind to HUB targets. According to gene ontology biological process analysis and Kyoto encyclopedia of genes and genomes pathway enrichment analysis, the PI3K-Akt signaling pathway, the MAPK signaling pathway, or the cAMP signaling pathway play a key role in treating of asthma by JXOL. The HUB target-miRNA network showed that 6 miRNAs were recognized. In our study, we have revealed for the first time the unique components, multiple targets, and diverse pathways in JXOL that underlie its mechanism of action in treating asthma through miRNAs.

Phylogenetic analysis of PP2C proteins and interactive proteins analyze of BjuPP2C52 in Brassica juncea.

Brassica juncea var. tumida Tsen et Lee (Tumorous stem mustard) is an unique vegetable in China. Its enlarged tumorous stem was used as main raw material to produce pickle (Zhacai). In practice, early-bolting happens around 15% of planting area all year and inhibits its production. Here, about 209 PP2C proteins were identified through HMMER software and divided into 13 sub-families in B. juncea. BjuPP2C52 belongs to E sub-family, was up-regulated at reproductive growth stages and interacts with BjuFKF1, a key protein in regulating plant photoperiod flowering, in vitro and in vivo. To explore interactive proteins, BjuPP2C52 was used as bait, 12 potential interactive proteins were screened from yeast library, and they are BjuCOL3, BjuCOL5, BjuAP2, BjuAP2-1, BjuSVP-1, BjuFLC-2, BjuSKP1f, BjuA014572, BjuA008686, BjuO002119, BjuB036787 and BjuA019268. Further study verified that 10 out of the 12 screened proteins interacted with BjuPP2C52 in vivo. qRT-PCR was conducted to understand the expression pattern of those 10 interactive proteins in different tissues and development stages in B. juncea. The results showed that BjuCOL3, BjuCOL5, BjuB036787 and BjuA019268 were significantly up-regulated, while BjuA008686 and BjuO002119 were down-regulated in flowers compared with other four tissues. In developmental stages, BjuCOL5, BjuAP2, BjuAP2-1, BjuA014572, BjuB036787 and BjuA019268 were significantly up-regulated, while BjuSVP-1, BjuA008686 and BjuO002119 were down-regulated at reproductive stages. Based on the results, BjuCOL5, BjuAP2, BjuAP2-1, BjuSVP-1, BjuA014572, BjuB036787 and BjuA019268 may function in regulating flowering time in B. juncea.

Detection of Major SARS-CoV-2 Variants of Concern in Clinical Samples via CRISPR-Cas12a-Mediated Mutation-Specific Assay.

Objectives: Emerging severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants pose a great threat and burden to global public health. Here, we evaluated a clustered regularly interspaced short palindromic repeat-associated enzyme 12a (CRISPR-Cas12a)-based method for detecting major SARS-CoV-2 variants of concern (VOCs) in SARS-CoV-2 positive clinical samples. Methods: Allele-specific CRISPR RNAs (crRNAs) targeting the signature mutations in the spike protein of SARS-CoV-2 are designed. A total of 59 SARS-CoV-2 positive oropharyngeal swab specimens were used to evaluate the performance of the CRISPR-Cas12a-mediated assay to identify major SARS-CoV-2 VOCs. Results: Compared with Sanger sequencing, the eight allele-specific crRNAs analyzed can specifically identify the corresponding mutations with a positive predictive value of 83.3-100% and a negative predictive value of 85.7-100%. Our CRISPR-Cas12a-mediated assay distinguished wild-type and four major VOCs (Alpha, Beta, Delta, and Omicron) of SARS-CoV-2 with a sensitivity of 93.8-100.0% and a specificity of 100.0%. The two methods showed a concordance of 98.3% (58/59) with a κ value of 0.956-1.000, while seven (11.9%) samples were found to be positive for extra mutations by the CRISPR-based assay. Furthermore, neither virus titers nor the sequences adjacent to the signature mutations were associated with the variation of fluorescence intensity detected or the false-positive reaction observed when testing clinical samples. In addition, there was no cross-reaction observed when detecting 33 SARS-CoV-2 negative clinical samples infected with common respiratory pathogens. Conclusions: The CRISPR-Cas12a-based genotyping assay is highly sensitive and specific when detecting both the SARS-CoV-2 wild-type strain and major VOCs. It is a simple and rapid assay that can monitor and track the circulating SARS-CoV-2 variants and the dynamics of the coronavirus disease 2019 (COVID-19) pandemic and can be easily implemented in resource-limited settings.

[Etiology survey on virus of acute respiratory infection in Guangzhou from 2006 to 2009].

OBJECTIVE: To investigate the pathogens of acute respiratory infection (ARI) in Guangzhou from 2006 to 2009. METHODS: A total of 1554 cases of ARI patients in Second Affiliated Hospital of Guangzhou Medical College from September 2006 to September 2009, were recruited in the survey. The sample of throat and pharyngeal swab were collected from each patient.11 types of virus including influenza A (FluA), influenza B (FluB), adenovirus (ADV), human rhinovirus (HRV), respiratory syncytial virus (RSV), parainfluenza virus type 1, type 2, type 3 (HPIV1, HPIV2, HPIV3), human metapneumovirus (MPV) and human coronavirus (HCoV) type 229E, type OC43 were detected by Fluorescence Quota PCR method. The epidemic feature and clinical characteristic of each virus were then analyzed. RESULTS: Virus were found in 1024 samples in total, accounting for 65.9% (1024/1554). RSV was the most common virus, which was found in 261 samples (16.8%); and followed by HRV as 13.9% (216/1554), FluA as 11.6% (181/1554), MPV as 6.5% (101/1554), FluB as 6.4% (99/1554), HPIV as 4.9% (76/1554), ADV as 3.5% (55/1554) and HCoV as 2.3% (35/1554). HPIV and HCoV shared a similar infection ratio among different age groups. The infection ratio of FluA and FluB was highest among 15-24 years old group, accounting for 16.5% (29/176) and 7.4% (13/176) respectively. MPV, RSV and HRV were the main pathogens caused infection among children under 4 years old, accounting for 9.7% (49/503), 21.7% (109/503) and 18.9% (95/503). The infection ratio of ADV was 6.0% (19/318), which was the most common pathogen among 5-14 years old patients. The incidence rate of HPIV and HRV showed no obvious seasonal features; while the prevalence of FluA, FluB, RSV, ADV, MPV and HCoV changed significantly in different seasons.22.2% (227/1024) ARI patients co-infected other respiratory virus.90.1% (163/181) FluA patients, 88.9% (88/99) FluB patients and 92.7% (51/55) ADV patients had high fever symptoms. CONCLUSION: RSV was the main pathogen of ARI, and the new-found virus MPV was also another crucial pathogen. Some pathogens' incidence rate were related to the season and patient's age. Co-infections of other respiratory virus were also detected in parts of ARI patients.

Analysis of differentially expressed genes and pathways associated with male sterility lines in watermelon via bulked segregant RNA-seq.

Genic male sterility (GMS) is a common and important trait, which is widely used for the production of hybrid seeds. However, the molecular mechanism of GMS in watermelon remains poorly understood. In this study, we comparatively analyzed the transcriptome profiles of sterile and fertile floral buds using the bulked segregant analysis (BSA) and transcriptome sequencing (RNA-seq). A total of 2507 differentially expressed genes (DEGs) including 593 up-regulated and 1914 down-regulated, were identified to be related to male sterility in watermelon line Se18. Gene ontology (GO) analysis showed that 57 GO terms were significantly enriched, while Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis revealed plant hormone signal transduction, glycolysis/gluconeogenesis, starch and sucrose metabolism, plant-pathogen interaction, phenylpropanoid biosynthesis pathways were obviously enriched. Furthermore, the efficiency of the RNA-seq analysis was validated by quantitative real-time PCR (qRT-PCR). Among the DEGs, some valuable candidate genes involved in pollen development were identified, such as gene Cla000029, a bHLH transcription factor and homologous to MS1 in Arabidopsis. Moreover, other DEGs including MYB gene Cla012590 (MYB26), Cla017100 (MYB21), etc., also provide useful information for further understanding the function of key genes involved in pollen development. This study provides new insights into the global network of male sterility in watermelon.

[Investigation of human metapneumovirus in children with acute respiratory tract infections in Guangzhou areas].

OBJECTIVE: To find out the status of human metapneumovirus (hMPV) infection in children under 14 years old with acute respiratory tract infections (ARI) in Guangzhou, analyze the epidemiology and clinical characteristics among the hMPV-infected children, and provide some basis for research of hMPV. METHODS: All 521 throat and pharyngeal swabs were collected among the children with acute respiratory tract infections in outpatient departments and those admitted to the wards from September 2006 to August 2008. Then total nucleic acid was extracted from respiratory specimens. The 213 nucleosides of nucleoprotein gene were detected by RT-PCR and 16 strong positive samples were picked to compare with the sequence of hMPV in GenBank after the sequence of the amplification products were determined. Then applied statistical analysis to the data of the collected patients. RESULTS: All 521 samples were detected by RT-PCR, and confirmed that N gene was positive in 39 samples with a detection rate of 7.49%, and the peak time was in October and April. The 16 amplification products were compared by using the analysis of gene sequence. The nucleocapsid protein (N) gene similarity to BJ1897 of Beijing was up to 99%, and to AY550156 of Thailand was up to 97%, genotype B was the most common genotype. CONCLUSION: There existed hMPV infection in children acute respiratory system diseases in Guangzhou areas, in which the children under the age of 6 years were accounted for the main group, however there was no difference in gender. The main symptoms of the patients with hMPV infection were high fever and cough symptom of catarrh. Co-infections other than respiratory virus with hMPV were detected as 41.03% of positive samples.

Transcriptomic and physiological analyses reveal drought adaptation strategies in drought-tolerant and -susceptible watermelon genotypes.

Drought stress has become one of the most urgent environmental hazards for horticultural crops. In this research, we analyzed watermelon adaptation strategies to drought stress in drought-tolerant (M20) and -susceptible (Y34) genotypes via transcriptomic and physiological analyses. After drought stress, a total of 6228 and 4311 differentially expressed genes (DEGs) were observed in Y34 and M20, respectively. Numerous DEGs were involved in various defense responses such as antioxidation, protein protection, osmotic adjustment, wax accumulation, hormone signaling, and melatonin biosynthesis. Accordingly, the contents of ABA, melatonin, wax, and some osmoprotectants were increased by drought stress in both Y34 and especially M20. Exogenous application of melatonin or ABA induced wax accumulation and drought tolerance and melatonin may function upstream of ABA. In comparison to Y34, M20 was more able to activate ABA signaling, melatonin biosynthesis, osmotic adjustment, antioxidation, and wax accumulation under drought stress. These stronger responses confer M20 tolerance to drought. Photosynthesis and most DEGs involved in photosynthesis and cell growth were decreased by drought stress in both M20 and especially Y34. For drought-susceptible genotypes, growth retardation may be an important mechanism for saving and redistributing resources in order to reprogram stress signaling networks.

Multiple outbreaks of acute hemorrhagic conjunctivitis due to a variant of coxsackievirus A24: Guangdong, China, 2007.

Acute hemorrhagic conjunctivitis (AHC) is usually caused by enterovirus 70, coxsackievirus A24(CA24v) and adenoviruses. Several outbreaks of AHC caused by a CA24v have occurred since it was imported into China in 1971. Multiple outbreaks of AHC reappeared in 10 cities of Guangdong during June to November in 2007. The epidemic began in the June, and spread extensively, with a peak in the September. A total of 31,659 cases were reported to center for disease control and prevention of Guangdong, it was estimated that the number of actual AHC was >200 thousands. Forty conjunctival swab specimens were collected from the cases diagnosed clinically with AHC. (RT)-PCR testing on these conjunctival specimens revealed the presence of an enterovirus, and this was confirmed by 16 isolates. We demonstrated the most likely etiological agent for the multiple outbreaks was a variant of coxsackievirus A24 by molecular typing using a partial VP1 sequence. Sequence comparison and phylogenetic analyses of the VP1 and 3Cpro gene regions were performed by Neighbor-joining method, the strains from different outbreaks and different geographical areas within Guangdong had no sequence divergence in 2007. The representative isolates from mainland of China including Hangzhou, Ningbo, Beijing, Yunnan, Liaoning, and Henan were analyzed in this study. Phylogenetic analysis revealed theses isolates were located in different clusters, a close phylogenetic and chronological relationship with Singaporean, South Korean and Thailand isolates had been observed. This confirms CA24v circulated in China's mainland has not evolved independently, but co-evolved with the isolates of Southeast Asia.

Identification and expression analyses of WRKY genes reveal their involvement in growth and abiotic stress response in watermelon (Citrullus lanatus).

Despite identification of WRKY family genes in numerous plant species, a little is known about WRKY genes in watermelon, one of the most economically important fruit crops around the world. Here, we identified a total of 63 putative WRKY genes in watermelon and classified them into three major groups (I-III) and five subgroups (IIa-IIe) in group II. The structure analysis indicated that ClWRKYs with different WRKY domains or motifs may play different roles by regulating respective target genes. The expressions of ClWRKYs in different tissues indicate that they are involved in various tissue growth and development. Furthermore, the diverse responses of ClWRKYs to drought, salt, or cold stress suggest that they positively or negatively affect plant tolerance to various abiotic stresses. In addition, the altered expression patterns of ClWRKYs in response to phytohormones such as, ABA, SA, MeJA, and ETH, imply the occurrence of complex cross-talks between ClWRKYs and plant hormone signals in regulating plant physiological and biological processes. Taken together, our findings provide valuable clues to further explore the function and regulatory mechanisms of ClWRKY genes in watermelon growth, development, and adaption to environmental stresses.

Genome-wide identification and expression analysis of calcium­dependent protein kinase and its related kinase gene families in melon (Cucumis melo L.).

The calcium-dependent protein kinase (CDPK) is a ser/thr protein kinase that plays vital roles in plant growth, development, and responses to multiple stresses. Despite an important member of the stress responsive gene family, little is known about the evolutionary history and expression patterns of CDPK genes in melon. Herein, a total of 18 CDPK genes and 7 CDPK-related protein kinases (CRK) genes were identified in the melon genome via bioinformatic analysis, which were unevenly distributed across eleven chromosomes with an apparent exception for chromosome 3. Comparative syntenic analysis between Cucumis melo L. and Arabidopsis thaliana revealed that 13 CmCDPKs and 19 AtCPKs existed in 20 corresponding syntenic blocks. In addition, based on gene structure and phylogenetic analyses, all CmCDPKs were divided into four groups (CDPK I-IV) and CmCRKs clustered into one group (CRK I). Interestingly, group CDPK IV was clearly distinct from the other three CDPK groups, but clustered with CRK I on the phylogenetic tree, implying their origination from a common ancestor. Furthermore, CmCDPKand CmCRK genes were differentially expressed in response to various stimuli, such as biotic stress (Podosphaera xanthii), abiotic stress (salt and cold), and hormone (abscisic acid) treatment. To our knowledge, this is the first report on CDPK and CRK gene families in melon, which provides a basic foundation for functional characterizations of CmCDPK and CmCRK genes in the future.

The Effects of Cattle Manure and Garlic Rotation on Soil under Continuous Cropping of Watermelon (Citrullus lanatus L.).

Continuous cropping of watermelon (Citrullus lanatus L.) can lead to reduced yield and quality. We aimed to determine the effects of cattle manure addition and rotation with green garlic to improve yield and reduce disease incidence in watermelon and to examine the effects on the biological and chemical characteristics of the soil. Field experiments were performed during 2012-2014 on land previously under two years of continuous watermelon cropping in northwest China. We examined three treatment combinations: watermelon and garlic rotation, cattle manure application before watermelon planting, and combined cattle manure addition and crop rotation. Watermelon monoculture was retained as a control. Watermelon yield was significantly higher and disease incidence was lower in the treatments than the control. The populations of soil bacteria and actinomycetes and the bacteria/fungi ratio increased significantly and soil enzyme activities were generally enhanced under treatments. Available nutrients and soil organic matter contents were much higher under experimental treatments than the control. Results suggest both cattle manure application and garlic rotation can ameliorate the negative effects of continuous cropping. The combined treatment of cattle manure addition and green garlic rotation was optimal to increase yield, reduce disease incidence and enhance soil quality.

Transcriptome Profiling of Watermelon Root in Response to Short-Term Osmotic Stress.

Osmotic stress adversely affects the growth, fruit quality and yield of watermelon (Citrullus lanatus (Thunb.) Matsum. & Nakai). Increasing the tolerance of watermelon to osmotic stress caused by factors such as high salt and water deficit is an effective way to improve crop survival in osmotic stress environments. Roots are important organs in water absorption and are involved in the initial response to osmosis stress; however, few studies have examined the underlying mechanism of tolerance to osmotic stress in watermelon roots. For better understanding of this mechanism, the inbred watermelon accession M08, which exhibits relatively high tolerance to water deficits, was treated with 20% polyethylene glycol (PEG) 6000. The root samples were harvested at 6 h after PEG treatment and untreated samples were used as controls. Transcriptome analyses were carried out by Illumina RNA sequencing. A total of 5246 differentially expressed genes were identified. Gene ontology enrichment and biochemical pathway analyses of these 5246 genes showed that short-term osmotic stress affected osmotic adjustment, signal transduction, hormone responses, cell division, cell cycle and ribosome, and M08 may repress root growth to adapt osmotic stress. The results of this study describe the watermelon root transcriptome under osmotic stress and propose new insight into watermelon root responses to osmotic stress at the transcriptome level. Accordingly, these results allow us to better understand the molecular mechanisms of watermelon in response to drought stress and will facilitate watermelon breeding projects to improve drought tolerance.

Regulation of Plant Growth, Photosynthesis, Antioxidation and Osmosis by an Arbuscular Mycorrhizal Fungus in Watermelon Seedlings under Well-Watered and Drought Conditions.

Drought stress has become an increasingly serious environmental issue that influences the growth and production of watermelon. Previous studies found that arbuscular mycorrhizal (AM) colonization improved the fruit yield and water use efficiency (WUE) of watermelon grown under water stress; however, the exact mechanisms remain unknown. In this study, the effects of Glomus versiforme symbiosis on the growth, physio-biochemical attributes, and stress-responsive gene expressions of watermelon seedlings grown under well-watered and drought conditions were investigated. The results showed that AM colonization did not significantly influence the shoot growth of watermelon seedlings under well-watered conditions but did promote root development irrespective of water treatment. Drought stress decreased the leaf relative water content and chlorophyll concentration, but to a lesser extent in the AM plants. Compared with the non-mycorrhizal seedlings, mycorrhizal plants had higher non-photochemical quenching values, which reduced the chloroplast ultrastructural damage in the mesophyll cells and thus maintained higher photosynthetic efficiency. Moreover, AM inoculation led to significant enhancements in the enzyme activities and gene expressions of superoxide dismutase, catalase, ascorbate peroxidase, glutathione reductase, and monodehydroascorbate reductase in watermelon leaves upon drought imposition. Consequently, AM plants exhibited lower accumulation of MDA, H2O2 and [Formula: see text] compared with non-mycorrhizal plants. Under drought stress, the soluble sugar and proline contents were significantly increased, and further enhancements were observed by pre-treating the drought-stressed plants with AM. Taken together, our findings indicate that mycorrhizal colonization enhances watermelon drought tolerance through a stronger root system, greater protection of photosynthetic apparatus, a more efficient antioxidant system and improved osmoregulation. This study contributes to advances in the knowledge of AM-induced drought tolerance.

Whole genomic sequence and replication kinetics of a new enterovirus C96 isolated from Guangdong, China with a different cell tropism.

Enterovirus 96 (EV-C96) is a newly described serotype within the enterovirus C (EV-C) species, and its biological and pathological characters are largely unknown. In this study, we sequenced the whole genome of a novel EV-C96 strain that was isolated in 2011 from a patient with acute flaccid paralysis (AFP) in Guangdong province, China and characterized the properties of its infection. Sequence analysis revealed the close relationship between the EV-C96 strains isolated from the Guangdong and Shandong provinces of China, and suggested that recombination events occurred both between these EV-C96 strains and with other EV-C viruses. Moreover, the virus replication kinetics showed EV-C96 Guangdong strain replicated at a high rate in RD cells and presented a different cell tropism to other strains isolated from Shandong recently. These findings gave further insight into the evolutionary processes and extensive biodiversity of EV-C96.

[Comparison of two nucleic acid extraction methods for norovirus in oysters].

OBJECTIVE: To explore a convenient and effective method for norovirus nucleic acid extraction from oysters suitable for long-term viral surveillance. METHODS: Two methods, namely method A (glycine washing and polyethylene glycol precipitation of the virus followed by silica gel centrifugal column) and method B (protease K digestion followed by application of paramagnetic silicon) were compared for their performance in norovirus nucleic acid extraction from oysters. Real-time RT-PCR was used to detect norovirus in naturally infected oysters and in oysters with induced infection. RESULTS: The two methods yielded comparable positive detection rates for the samples, but the recovery rate of the virus was higher with method B than with method A. CONCLUSION: Method B is a more convenient and rapid method for norovirus nucleic acid extraction from oysters and suitable for long-term surveillance of norovirus.

[Epidemiological and etiological characteristics of diarrheal disease among children under 5 years of age in Guangdong province, in 2012].

OBJECTIVE: To analyze the epidemiological and etiological characteristics of diarrheal disease among children under 5 years of age in Guangdong province, in 2012. METHODS: 64 hospitals in 21 cities were chosen as the diarrheal syndromic surveillance sites, of which 14 hospitals were selected to carry out etiological surveillance among children under 5 years of age, including isolation and culture of both Vibrio cholera and Shigella as well as nucleic acid detection of rotavirus and norovirus by PCR. Descriptive method was used to analyze data from syndromic and etiological surveillance programs on diarrheal, from 1932 parents of the children. RESULTS: In 2012, the outpatient attendance rate on diarrheal among children under 5 years was 0.8%. The proportion of diarrheal in children under 5-year-olds was 63.5%, among the total number of diarrheal outpatients at the outpatient clinics under surveillance program. The morbidity of infectious diarrhea was 1454.5/10 million in children under 5 years of age. A total number of 1932 specimens were collected from children under 5 years of age, in the outpatient department. Among these specimens,Vibrio cholera appeared all negative but one was Shigella positive and proved to be Sh. sonnei. The positive rates of rotavirus and norovirus were 14.1% (273/1932)and 16.9% (326/1932). Both rotavirus and norovirus were found in 24 specimens, with a positive rate as 1.2% . 112 specimens were successfully gene sequenced for rotavirus, of which 33.9% as G1[P8] genotype, 25.9% as G9[P8], 12.5% as G2[P4] and 9.8% as G3[P8] respectively. 90 specimens were successfully gene-sequenced for norovirus, of which 76.7% as G II.4 genotype. Genetic subtypes of G II. 4/2006b, accounted for 50.0% and could be detected around the year except for June and December. New G II. 4/Sydney Strain_2012 was first detected in August and became the predominant in December. In addition, 5 specimens belonged to G I genotype with other 16 subtypes of G II. CONCLUSION: Results from our study proved that children under 5 years of age belonged to high-risk group for diarrheal disease in Guangdong province. Rotavirus and norovirus were both diverse in terms of genome.